Molecular analysis of hepatitis B virus (HBV) in an HIV co-infected patient with reactivation of occult HBV infection following discontinuation of lamivudine-including antiretroviral therapy

Abstract Background Occult hepatitis B virus (HBV) infection (OBI) is characterized by HBV DNA persistence even though the pattern of serological markers indicates an otherwise resolved HBV infection. Although OBI is usually clinically silent, immunocompromised patients may experience reactivation of the liver disease. Case presentation We report the case of an individual with human immunodeficiency virus (HIV) infection and anti-HBV core antibody positivity, who experienced severe HBV reactivation after discontinuation of lamivudine-including antiretroviral therapy (ART). HBV sequencing analysis showed a hepatitis B surface antigen escape mutant whose presence in an earlier sample excluded reinfection. Molecular sequencing showed some differences between two isolates collected at a 9-year interval, indicating HBV evolution. Resumption of ART containing an emtricitabine/tenofovir combination allowed control of plasma HBV DNA, which fell to undetectable levels. Conclusion This case stresses the ability of HBV to evolve continuously, even during occult infection, and the effectiveness of ART in controlling OBI reactivation in HIV-infected individuals.</p

Human cell types important for Hepatitis C Virus replication in vivo and in vitro. Old assertions and current evidence

Hepatitis C Virus (HCV) is a single stranded RNA virus which produces negative strand RNA as a replicative intermediate. We analyzed 75 RT-PCR studies that tested for negative strand HCV RNA in liver and other human tissues. 85% of the studies that investigated extrahepatic replication of HCV found one or more samples positive for replicative RNA. Studies using in situ hybridization, immunofluorescence, immunohistochemistry, and quasispecies analysis also demonstrated the presence of replicating HCV in various extrahepatic human tissues, and provide evidence that HCV replicates in macrophages, B cells, T cells, and other extrahepatic tissues. We also analyzed both short term and long term in vitro systems used to culture HCV. These systems vary in their purposes and methods, but long term culturing of HCV in B cells, T cells, and other cell types has been used to analyze replication. It is therefore now possible to study HIV-HCV co-infections and HCV replication in vitro

GB individuals virus C replication in cerebrospinal fluid of HIV positive

Background GB virus C (GBV-C) is an orphan Flavivirus distantly related to hepatitis C virus (HCV). GBV-C infection in humans is common and its transmission is similar to both HCV and HIV, including sexual, parenteral and vertical transmission. The aim of this study was to investigate the presence and compartimentalization of GBV-C strains in plasma, PBMC and cerebrospinal fluid (CSF) of HIV positive patients in an attempt to identify GBV-C replication in CSF. Methods This retrospective study involved 22 HIV positive patients who underwent a lumbar puncture for diagnostic purposes. To evaluate the prevalence of GBV-C infection in our Centre, a control group of 150 HIV positive patients was included in the study. GBV-C-RNA was searched for in paired plasma, CSF and PBMC by means of nested PCR for 5\u2019UTR. GBV-C genotype was identified by direct sequencing and phylogenetical analysis. GBV-C population in plasma and CSF was characterized by analysis of at least 25 clones for each compartment. HIV load was measured by a quantitative PCR (Amplicor HIV Monitor). Results GBV-C-RNA was detected in plasma from 37/150 (25%) control patients and in the PBMC from 3/34 (9%) patients with GBV-C detected in plasma. GBV-C-RNA was identified in 5/22 (23%) plasma, in none of PBMC and in 2/5 (40%) CSF samples obtained from the 5 GBV-C positive patients in plasma. This data showed that the presence of GBV-C-RNA in plasma was similar in control group and study population. Albeit GBV-C-RNA was detected in 3 PBMC of control group and in none PBMC of the study population, the difference was not statistically significant. Direct sequencing of GBV-C 5\u2019UTR detected in plasma revealed the presence of genotype 2 in three patients GBV-C-RNA negative in CSF. Analysis of GBV-C population within the 5\u2019UTR showed the presence of GBV-C genotype 2 in plasma and genotype 3 in CSF of one patient, whereas the other case had mixed infection in plasma (genotype 3/1) and genotype 3 alone in CSF. Plasma HIV-RNA levels were significantly higher in patients with CSF positive for GBV-C than in those with GBV-C negative CSF (mean values 5.35 vs 4.39 Log copies/ml, p=0.027). HIV viral load in CSF was similar in all patients (mean values 3.66 vs 3.14 Log copies/ml). Conclusion These findings suggest the replication of GBV-C in CSF and the selection of genotype 3 in this compartment. The presence of discordant genotypes in paired plasma/CSF of one patient could be due to a past mixed infection in plasma

REPLICAZIONE DEL GB-C VIRUS NEL LIQUIDO CEREBROSPINALE DI INDIVIDUI HIV POSITIVI

Premessa e scopo dello studio Il GB-C virus (GBV-C) \ue8 un Flavivirus orfano, filogeneticamente correlato al virus dell\u2019epatite C (HCV). Le vie di trasmissione sono le medesime di HIV e HCV: sessuale, parenterale e verticale. Questo studio si propone di analizzare la presenza e la compartimentalizzazione delle varianti di GBV-C nel plasma, nei PBMC e nel liquido cerebrospinale (LCS) di pazienti HIV positivi. Pazienti e metodi Sono stati inclusi nello studio 22 pazienti HIV positivi sottoposti a puntura lombare a scopo diagnostico. Un gruppo di 150 pazienti sieropositivi in cura presso il nostro Centro \ue8 stato incluso come controllo. La presenza di GBV-C \ue8 stata ricercata nei campioni accoppiati di plasma, PBMC e LCS mediante una nested PCR specifica per la regione 5\u2019UTR. Il genotipo di GBV-C \ue8 stato identificato mediante sequenziamento diretto e analisi filogenetica. La popolazione virale \ue8 stata studiata, all\u2019interno di ciascun compartimento, mediante l\u2019analisi di almeno 25 cloni. Risultati Nel gruppo di controllo il GBV-C-RNA \ue8 stato rilevato nel plasma di 37/150 (25%) pazienti e nei PBMC di 3/34 (9%) individui risultati GBV-C positivi nel plasma. Nella popolazione analizzata, il GBV-C-RNA \ue8 stato identificato in 5/22 (23%) campioni di plasma, in nessun PBMC e in 2/5 (40%) campioni di LCS ottenuti dai 5 pazienti risultati positivi nel plasma. La prevalenza dell\u2019infezione da GBV-C nel plasma risulta quindi simile nella popolazione studiata e nel gruppo di controllo. Nonostante il GBV-C-RNA sia stato evidenziato in 3 PBMC ottenuti dal gruppo di controllo e in nessuno di quelli isolati dalla popolazione analizzata, la differenza non risulta statisticamente significativa. Il sequenziamento diretto della regione 5\u2019UTR del GBV-C ha rivelato la presenza del genotipo 2 nel plasma degli individui negativi nel LCS. L\u2019analisi della popolazione virale nei 2 pazienti LCS positivi, ha mostrato, in un caso il genotipo 2 e 3 rispettivamente in plasma e LCS e nell\u2019altro un\u2019infezione mista nel plasma (genotipo 3/1) e solo il genotipo 3 nel LCS. La viremia HIV plasmatica risulta significativamente pi\uf9 elevata nei pazienti GBV-C positivi rispetto a quelli GBV-C negativi nel LCS (viremia media: 5.35 vs 4.39 Log copie/ml, p=0.027). Tutti i pazienti mostrano livelli di HIV-RNA simili nel LCS ( media: 3.66 vs 3.14 Log copie/ml). Conclusioni Questi risultati suggeriscono che GBV-C sia in grado di replicare nel LCS e che il genotipo 3 possa essere selezionato in questo compartimento

Hepatic safety profile of darunavir with low dose ritonavir (DRV/r) in HIV/HCV coinfected and HIV monoinfected patients.

The hepatic safety profile of ART including DRV/r was retrospectively evaluated in antiretroviral-experienced HIV-infected patients (18 HIV/HCV coinfected, group A and 29 infected with HIV alone, group B) during a 72 week study. During the study, liver enzyme values were higher in group A, but in the case of abnormal transaminase levels, the median values did not exceed 1.6xULN. This study showed evidence of long-lasting hepatic safety of ART including PI DRV/r in HIV/HCV coinfected and in HIV monoinfected persons